{"title":"下一代自动膜片钳仪器QPatch II对GABAA受体电生理的研究","authors":"Melanie Schupp, Sung Hoon Park, Binbin Qian, Weifeng Yu","doi":"10.1002/cpph.75","DOIUrl":null,"url":null,"abstract":"<p>Ligand-gated ion channels (LGICs) are a group of diverse ion channels that are gated by ligands and play important roles in normal physiological and pathological conditions. Many of them are drug targets that have been pursued, are being pursued, and will likely be pursued in the future by pharmaceutical companies and academic groups for a variety of diseases. One of those LGICs is the GABA<sub>A</sub> receptor, a heterooligomeric chloride channel that can be blocked and modulated at various sites. In order to study the receptor's functional response to compounds, the manual patch-clamp method provides a detailed but low-throughput electrophysiological characterization. QPatch II, a next-generation automated patch clamp machine that was recently developed by Sophion Bioscience, provides an automated electrophysiological study of ion channels. In this article, we use the GABA<sub>A</sub> receptor as an example for studying LGICs and describe two detailed protocols for using QPatch II to carry out pharmacological studies on the receptor. © 2020 by John Wiley & Sons, Inc.</p><p><b>Basic Protocol 1</b>: Ligand concentration-response experiment (GABA<sub>A</sub> receptor) on QPatch II</p><p><b>Alternate Protocol</b>: Non-cumulative ligand concentration-response experiment (GABA<sub>A</sub> receptor) on QPatch II</p><p><b>Support Protocol 1</b>: Cell culture of HEK293-hGABA<sub>A</sub> (α5β3γ2)</p><p><b>Support Protocol 2</b>: Data analysis for Basic Protocol 1</p><p><b>Support Protocol 3</b>: Data analysis for Alternate Protocol</p><p><b>Basic Protocol 2</b>: Antagonist dose-response experiment (GABA<sub>A</sub> receptor) on QPatch II</p><p><b>Support Protocol 3</b>: Data analysis for Basic Protocol 2</p>","PeriodicalId":10871,"journal":{"name":"Current Protocols in Pharmacology","volume":"89 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpph.75","citationCount":"7","resultStr":"{\"title\":\"Electrophysiological Studies of GABAA Receptors Using QPatch II, the Next Generation of Automated Patch-Clamp Instruments\",\"authors\":\"Melanie Schupp, Sung Hoon Park, Binbin Qian, Weifeng Yu\",\"doi\":\"10.1002/cpph.75\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Ligand-gated ion channels (LGICs) are a group of diverse ion channels that are gated by ligands and play important roles in normal physiological and pathological conditions. Many of them are drug targets that have been pursued, are being pursued, and will likely be pursued in the future by pharmaceutical companies and academic groups for a variety of diseases. One of those LGICs is the GABA<sub>A</sub> receptor, a heterooligomeric chloride channel that can be blocked and modulated at various sites. In order to study the receptor's functional response to compounds, the manual patch-clamp method provides a detailed but low-throughput electrophysiological characterization. QPatch II, a next-generation automated patch clamp machine that was recently developed by Sophion Bioscience, provides an automated electrophysiological study of ion channels. In this article, we use the GABA<sub>A</sub> receptor as an example for studying LGICs and describe two detailed protocols for using QPatch II to carry out pharmacological studies on the receptor. © 2020 by John Wiley & Sons, Inc.</p><p><b>Basic Protocol 1</b>: Ligand concentration-response experiment (GABA<sub>A</sub> receptor) on QPatch II</p><p><b>Alternate Protocol</b>: Non-cumulative ligand concentration-response experiment (GABA<sub>A</sub> receptor) on QPatch II</p><p><b>Support Protocol 1</b>: Cell culture of HEK293-hGABA<sub>A</sub> (α5β3γ2)</p><p><b>Support Protocol 2</b>: Data analysis for Basic Protocol 1</p><p><b>Support Protocol 3</b>: Data analysis for Alternate Protocol</p><p><b>Basic Protocol 2</b>: Antagonist dose-response experiment (GABA<sub>A</sub> receptor) on QPatch II</p><p><b>Support Protocol 3</b>: Data analysis for Basic Protocol 2</p>\",\"PeriodicalId\":10871,\"journal\":{\"name\":\"Current Protocols in Pharmacology\",\"volume\":\"89 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-04-09\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/cpph.75\",\"citationCount\":\"7\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Protocols in Pharmacology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpph.75\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"Pharmacology, Toxicology and Pharmaceutics\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Pharmacology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpph.75","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Pharmacology, Toxicology and Pharmaceutics","Score":null,"Total":0}
引用次数: 7
Electrophysiological Studies of GABAA Receptors Using QPatch II, the Next Generation of Automated Patch-Clamp Instruments
Ligand-gated ion channels (LGICs) are a group of diverse ion channels that are gated by ligands and play important roles in normal physiological and pathological conditions. Many of them are drug targets that have been pursued, are being pursued, and will likely be pursued in the future by pharmaceutical companies and academic groups for a variety of diseases. One of those LGICs is the GABAA receptor, a heterooligomeric chloride channel that can be blocked and modulated at various sites. In order to study the receptor's functional response to compounds, the manual patch-clamp method provides a detailed but low-throughput electrophysiological characterization. QPatch II, a next-generation automated patch clamp machine that was recently developed by Sophion Bioscience, provides an automated electrophysiological study of ion channels. In this article, we use the GABAA receptor as an example for studying LGICs and describe two detailed protocols for using QPatch II to carry out pharmacological studies on the receptor. © 2020 by John Wiley & Sons, Inc.
Basic Protocol 1: Ligand concentration-response experiment (GABAA receptor) on QPatch II
Alternate Protocol: Non-cumulative ligand concentration-response experiment (GABAA receptor) on QPatch II
Support Protocol 1: Cell culture of HEK293-hGABAA (α5β3γ2)
Support Protocol 2: Data analysis for Basic Protocol 1
Support Protocol 3: Data analysis for Alternate Protocol
Basic Protocol 2: Antagonist dose-response experiment (GABAA receptor) on QPatch II
Support Protocol 3: Data analysis for Basic Protocol 2
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